Safety Assessment of Eucalyptus globulus (Eucalyptus) - Derived Ingredients as Used in Cosmetics Status: Draft Report for Panel Review Release Date: November 10, 2017 Panel Meeting Date: December 4-5, 2017 The 2017 Cosmetic Ingredient Review Expert Panel members are: Chair, Wilma F. Bergfeld, M.D., F.A.C.P.; Donald V. Belsito, M.D.; Ronald A. Hill, Ph.D.; Curtis D. Klaassen, Ph.D.; Daniel C. Liebler, Ph.D.; James G. Marks, Jr., M.D.; Ronald C. Shank, Ph.D.; Thomas J. Slaga, Ph.D.; and Paul W. Snyder, D.V.M., Ph.D. The CIR Executive Director is Bart Heldreth, Ph.D. This report was prepared by Lillian C. Becker, Scientific Analyst/Writer. © Cosmetic Ingredient Review 1620 L Street, NW, Suite 1200 ♢ Washington, DC 20036-4702 ♢ ph 202.331.0651 ♢ fax 202.331.0088 [email protected] Commitment & Credibility since 1976 MEMORANDUM To: CIR Expert Panel and Liaisons From: Lillian C. Becker, M.S. Scientific Analyst and Writer Date: November 10, 2017 Subject: Safety Assessment of Eucalyptus globulus (Eucalyptus)-Derived Ingredients As Used In Cosmetics Attached is the draft report of Eucalyptus globulus (Eucalyptus)-derived ingredients as used in cosmetics. [Eucaly122017Rep] In September 2017, the Scientific Literature Review was posted for public comment with a request for additional data, including clarification of the definition of Eucalyptus Globulus Leaf Oil, which states that the oil may be sourced from other Eucalyptus species [Eucalyptus Globulus Leaf Oil is the volatile oil obtained from the leaves of Eucalyptus globulus and other species of Eucalyptus]. The source for all of these ingredients is the leaf or leaf/twig of the plant. The reported functions of the Eucalyptus globulus (eucalyptus)-derived ingredients include abrasive, fragrance ingredient, and skin-conditioning agent (miscellaneous and occlusive). A letter has been sent to RIFM asking their intentions towards the safety assessment of the fragrance-only ingredients recited in this report: Eucalyptus Globulus Leaf/Twig Oil and Eucalyptus Globulus Leaf Water. The Council has provided concentrations of use data and characterization of Eucalyptus Globulus Leaf Oil. [Eucaly122017Data_1-3] No other data have been provided. Council comments have been addressed. [Eucaly122017PCPC] In most cases, the main component of Eucalyptus Globulus Leaf Oil is reported to be eucalyptol (54% to 95%; also called 1,8-cineole or cineole). Eucalyptol is a cosmetic ingredient that has not been reviewed by CIR. Should this ingredient be added to this safety assessment? To help the Panel make this decision, a summary of a sampling of toxicity and relevant data with regard to eucalyptol are provided with this memo. If no further data are needed, the Panel should formulate a Discussion and issue a Tentative Report. However, if additional data are required, the Panel should be prepared to identify those needs and issue an Insufficient Data Announcement. Eucalyptol (CAS No. 470-82-6) Eucalyptol is the organic compound that conforms to the formula: (structure). It is the chief component of Eucalyptus Globulus Leaf Oil.1 2017 VCRP Data2 05F - Shampoos (non-coloring) EUCALYPTOL 2 05G - Tonics, Dressings, and Other Hair Grooming Aids EUCALYPTOL 1 08F - Nail Polish and Enamel Removers EUCALYPTOL 1 09B - Mouthwashes and Breath Fresheners EUCALYPTOL 12 12A - Cleansing EUCALYPTOL 3 12C - Face and Neck (exc shave) EUCALYPTOL 1 12J - Other Skin Care Preps EUCALYPTOL 2 Total 22 ABSORPTION, DISTRIBUTION, METABOLISM, AND EXCRETION Eucalyptol undergoes oxidation in vivo with the formation of hydroxycineole which is excreted as glucuronide.3 In rats, 2-hydroxycineole, 3-hydroxycineole and 1,8-dihydroxycineol-9-oic acid were identified as main urinary metabolites. After oral administration to brushtail possums (Trichosurus vulpecula), p-cresol, 9-hydroxycineole, and cineol-9-oic acid were found in urine. Rabbits given eucalyptol by gavage excreted 2-exo- and 2-endo-hydroxycineole as well as 3-exo- and 3-endo-hydroxycineole in the urine. Table 1. Overview of toxicity data on Eucalyptol. Assay Animal Concentration Results Reference Acute oral toxicity Rats LD50 = 2480 mg/kg 3 Acute oral toxicity Rats LD50 =1560 mg/kg 3 Lethal dose caused rapid cyanosis and stupor accompanied by irregular breathing, extreme sensitivity to noise, convulsions, and death from respiratory failure. Acute oral toxicity Mice 500 mg/kg An increase in liver enzyme activity was also found in 3 mice given 500 mg/kg orally. Table 1. Overview of toxicity data on Eucalyptol. Assay Animal Concentration Results Reference Subacute Oral Fischer 344 rats Stomach tube: 150, At dose levels of 600 mg/kg and higher, dose-related 3 Toxicity - 28 days (6/sex) 300, 600 and 1200 decrease of body weight gain and absence of a normal either by stomach mg/kg degree of hepatic centrilobular cytoplasmic vacuolization tube (5 days/week) Encapsulated form in was observed in male rats. Other dose-related lesions in or in encapsulated diet: 3750, 7500, the liver, kidneys and parotid salivary glands were found form with the diet. 15,000 and 30,000 at all dose levels in male rats fed encapsulated eucalyptol. mg/kg, equivalent to 381 to 3342 mg/kg/day for males and 353 to 3516 mg/kg/day for females. Subacute Oral Male Wistar rats 0, 500, or 1000 There were decreases in terminal body weight and 3 Toxicity – (10) mg/kg/day increased relative liver and kidney weights in both administered by treatment groups. The relative brain weight was increased gavage for 28 days in 1000 mg/kg/day group. No macroscopic changes were observed. Only brain, liver and kidneys were examined histopathologically. No changes in the brain were observed; minor focal infiltration of mononuclear cells in liver was observed in all groups. In kidneys, a dose- related accumulation of eosinophilic protein droplets containing α2u-globulin in the cytoplasm of proximal tubular epithelial cells was observed. Subacute Oral B6C3F1 mice Stomach tube: 150, The liver weight/body weight ratio in males was 3 Toxicity - 28 days (6/sex) 300, 600 and 1200 increased at all but the lowest dose given in encapsulated either by stomach mg/kg form as was the brain weight/body weight ratio in tube (5 days/week) Encapsulated form in females at the top dose level. Microscopic examination or in encapsulated diet: 3750, 7500, revealed a minimal hypertrophy of centrilobular form with the diet. 15000 and 30000 hepatocytes in mice of both sexes fed the encapsulated mg/kg, equivalent to compound, especially at the two highest dose levels. 600 to 5607 mg/kg/day for males and 705-6777 mg/kg/day for females. Chronic Oral Pathogen-free CFLP 0, 8 and 32 mg/kg/day No treatment-related effects on body weights, feed 3 Toxicity (and mice (52) in 1 mL toothpaste consumption, survival, weight of adrenals, kidneys, liver, Carcinogenicity) – base/kg/day lungs or spleen, on the microscopic appearance of brain, toothpaste lungs, liver and kidneys and on the tumor incidence were administered by observed. gavage (6 days/ week) for 80 weeks followed by 16 and 24 weeks rest. Genotoxicity – Salmonella Not specified No mutagenic effects with or without metabolic 3 Ames assay typhimurium (TA98, activation TA100, TA1535, and TA1537) Genotoxicity – S. typhimurium Not specified No mutagenic effects with or without metabolic 3 Ames assay (TA97a, TA98, activation TA100, and TA102) Genotoxicity – Chinese hamster Not specified No induced chromosome aberrations with or without 3 Chromosome ovary cells metabolic activation aberration assay Genotoxicity – Chinese hamster Not specified Sister chromatid exchanges were induced in CHO cells 3 Sister chromatid ovary cells only in the absence of metabolic activation at doses that exchange assay induced cell cycle delay. Genotoxicity – rec Bacillus subtilis Not specified No evidence of DNA damage 3 assay Genotoxicity – rec Bacillus subtilis Not specified No evidence of DNA damage 3 assay Dermal Irritation Human epidermis 100% Non-irritant. Relative mean viability of the treated tissue 4 In Vitro - Episkin™ model was 88.9% after 15 min exposure. 5 Dermal Irritation – Albino mice (10) Not specified Irritant dose in 50% of test individuals (ID50) = 1.008 Open mouse ear µg/5µL (0.0202%) assay Table 1. Overview of toxicity data on Eucalyptol. Assay Animal Concentration Results Reference Sensitization - local Female mice (5) 25 % and 50 % v/v in Stimulation Index (SI): 25%, 1.43; 50%, 2.03; 100%, 4 lymph node assay acetone/olive oil 4:1, 5.08. (LLNA) and 100 % v/v The concentration of Eucalyptol expected to cause a 3- fold increase in 3HTdR incorporation (EC3 value) was calculated to be 65.90%. Eucalyptol was considered to be a sensitizer under the conditions of the test. Ocular Irritation - Bovine cornea 100% Not considered to be an ocular corrosive or severe irritant 4 In Vitro - Bovine Corneal Opacity and Permeability Assay References 1. Nikitakis, J and Lange B (eds). Web-Based Ingredient Dictionary (wINCI). http://webdictionary.personalcarecouncil.org/jsp/Home.jsp. Washington, DC. Last Updated 2017. 2. Food and Drug Administration (FDA). Frequency of use of cosmetic ingredients; FDA Database. Washington, DC, FDA. 2017. 3. Scientific Committee on Food. Opinion of the Scientific Committee on Food on eucalyptol. Bruxelles/Brussel, Belgium, European Commission, Health & Comsumer Protection Directorate-General. 2002. https://ec.europa.eu/food/sites/food/files/safety/docs/sci- com_scf_out126_en.pdf. Report No. SCF/CS/FLAV/FLAVOUR/20 ADD2 FINAL. pp. 1-10. 4. European Chemicals Agency (ECHA). Cineole. European Chemicals